Transgenic Modifications of the Rat Genome

Abstract

The laboratory rat (R. norvegicus) is a very important experimental animal in several fields of biomedical research. This review describes the various techniques that have been used to generate transgenic rats: classical DNA microinjection and more recently described techniques such as lentiviral vector-mediated DNA transfer into early embryos, sperm-mediated transgenesis, embryo cloning by nuclear transfer and germline mutagenesis. It will also cover techniques associated to transgenesis such as sperm cryopreservation, embryo freezing and determination of zygosity. The availability of several technologies allowing genetic manipulation in the rat coupled to genomic data will allow biomedical research to fully benefit from the rat as an experimental animal.

Efficient gene targeting by homology-directed repair in rat zygotes using TALE nucleases

“…RI of the GFP expression cassette (scored as GFP expression and positive PCRs for GFP but with negative 59 and 39 junction PCRs) occurred at frequencies similar to those observed when linearized DNA is microinjected for the generation of transgenic rats (0.67%-2.54%) (Tesson et al 2005) and without differences among the different experimental conditions. These RI transgenic animals did not contain HDR events, and for Hprt1.2 TALE nucleases included animals 13.…”

Gene Therapy of Some Genetic Diseases by Transferring Normal Human Genomic DNA into Somatic Cells and Stem Cells from Patients

“…Microinjection technique has been used in transgenic animals for many years (Bishop & Smith, 1989;Chan & Yang, 2009;Charreau et al, 1996;Filipiak & Saunders, 2006;Ménoret et al, 2010;Tesson et al, 2005;Yang et al, 2008). Microinjection technique also has been used as a tool to clone animals-first, an unfertilized egg's nucleus is removed; then a nucleus of a somatic cell is microinjected into the denucleated egg; now the egg contains a whole copy of the diploid genomic DNA from the somatic cell and can be cultured in vitro to form a blastocyst; and the blastocyst is implanted into the womb of an animal; eventually a cloned animal is born (Campbell et al, 1996;Vajta & Gjerris, 2006;Willadsen, 1986;Wilmut et al, 1997).…”

20 Years of Hypertension Research using Genetically Modified Animals: No Clinically Promising Approaches in Sight

“…this is different from other literature searches, where basic research is usually cited more often (see for instance : lindl et al, 2005). the retrieved abstracts of original publications (except reviews) described animal models for hypertension (Denton et al, 2006;tesson et al, 2005;Kapoor and Dikshit, 2005), but contained no indications of new therapies for hypertension. A further, more recent search of PubMed for all articles from 2007 until April 2008 also did not show any clinically relevant publications.…”